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Complete de novo assembly of Wolbachia endosymbiont of Diaphorina citri Kuwayama (Hemiptera: Liviidae) using long-read genome sequencing.

ABSTRACT: Wolbachia, a gram-negative [Formula: see text]-proteobacterium, is an endosymbiont found in some arthropods and nematodes. Diaphorina citri Kuwayama, the vector of 'Candidatus Liberibacter asiaticus' (CLas), are naturally infected with a strain of Wolbachia (wDi), which has been shown to colocalize with the bacteria pathogens CLas, the pathogen associated with huanglongbing (HLB) disease of citrus. The relationship between wDi and CLas is poorly understood in part because the complete genome of wDi has not been available. Using high-quality long-read PacBio circular consensus sequences, we present the largest complete circular wDi genome among supergroup-B members. The assembled circular chromosome is 1.52 megabases with 95.7% genome completeness with contamination of 1.45%, as assessed by checkM. We identified Insertion Sequences (ISs) and prophage genes scattered throughout the genomes. The proteins were annotated using Pfam, eggNOG, and COG that assigned unique domains and functions. The wDi genome was compared with previously sequenced Wolbachia genomes using pangenome and phylogenetic analyses. The availability of a complete circular chromosome of wDi will facilitate understanding of its role within the insect vector, which may assist in developing tools for disease management. This information also provides a baseline for understanding phylogenetic relationships among Wolbachia of other insect vectors.

SUBMITTER: Neupane S

PROVIDER: S-EPMC8741817 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Diaphorina citri Kuwayama (Hemiptera: Liviidae), commonly known as Asian citrus psyllid (ACP), is an invasive insect pest and the vector of the bacterium causing Huanglongbing (HLB), a lethal disease of citrus. In the United States, ACP has been established in all citrus-producing zones, all of which have different environmental conditions. The spread of ACP and, more importantly, HLB, has progressed differently depending on the state, with more rapid spread in Florida and Texas than in California. Climatic variations between the regions are likely a strong factor in the difference in the rate of spread. Despite this, it is unknown how the flight capacity of D. citri is influenced by high temperatures (>30 °C) and subsequently, low humidity experienced in California but not in Texas or Florida. In this study, by using a custom-made, temperature-controlled flight mill arena, we assessed the effect of high temperatures on the flight capacity and flight propensity of D. citri under low (20-40%) and high (76-90%) relative humidity conditions. We found that temperature and humidity influence the propensity to engage in short or long-distance flight events. Psyllids exposed to temperatures above 43 °C only performed short flights (˂60 s), and a high relative humidity significantly decrease the proportion of long flights (≥60 s) at 26 and 40 °C. The flight capacity for insects who engaged in short and long flights was significantly affected by temperature but not by humidity. For long flyers, temperature (in the 26-43 °C range) was negatively correlated with distance flown and flight duration. The most favorable temperature for long dispersion was 26 °C, with suboptimal temperatures in the range of 32-37 °C and the least favorable temperatures at 40 and 43 °C. In conclusion, D. citri is able to fly in a broad range of temperatures and efficiently fly in high and low humidity. However, temperatures above 40 °C, similar to those experienced in semi-arid environments like Southern California or Arizona, are detrimental for its flight capacity.

Project description:The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is the most widespread and devastating pest species in citrus orchards and is the natural vector of the phloem-limited bacterium that causes Huanglongbing (HLB) disease. Thus, reducing the population of D. citri is an important means to prevent the spread of HLB disease. Due to the long-term use of chemical control, biological control has become the most promising strategy. In this study, a novel highly pathogenic fungal strain was isolated from naturally infected cadavers of adult D. citri. The species was identified as Aspergillus fijiensis using morphological identification and phylogenetic analysis and assigned the strain name GDIZM-1. Tests to detect aflatoxin B1 demonstrated that A. fijiensis GDIZM-1 is a non-aflatoxin B1 producer. The pathogenicity of the strain against D. citri was determined under laboratory and greenhouse conditions. The results of the laboratory study indicated that nymphs from the 1st to 5th instar and adults of D. citri were infected by A. fijiensis GDIZM-1. The mortality of nymphs and adults of D. citri caused by infection with A. fijiensis increased with the concentration of the conidial suspension and exposure time, and the median lethal concentration (LC50) and median lethal time (LT50) values gradually decreased. The mortality of D. citri for all instars was higher than 70%, with high pathogenicity at the 7th day post treatment with 1 × 108 conidia/mL. The results of the greenhouse pathogenicity tests showed that the survival of D. citri adults was 3.33% on the 14th day post-treatment with 1 × 108 conidia/mL, which was significantly lower than that after treatment with the Metarhizium anisopliae GDIZMMa-3 strain and sterile water. The results of the present study revealed that the isolate of A. fijiensis GDIZM-1 was effective against D. citri and it provides a basis for the development of a new microbial pesticide against D. citri after validation of these results in the field.

Dataset Information

Complete de novo assembly of Wolbachia endosymbiont of Diaphorina citri Kuwayama (Hemiptera: Liviidae) using long-read genome sequencing.

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